百日咳毒素通过诱导中性粒细胞胞外诱捕网形成参与实验性自身免疫性肌炎小鼠间质性肺疾病的发生  

Pertussis toxin in involves the development of interstitial lung disease in an experimental autoimmune myositis mice modelby inducing the formation of neutrophil extracellular traps

作　　者：白灵 马文兰 李飞飞 赵佩佩 朱嘉睿 张思功[3] Bai Ling;Ma Wenlan;Li Feifei;Zhao Peipei;Zhu Jiarui;Zhang Sigong(Second Clinical Medical College of Lanzhou University,Lanzhou 730030,China;Cuiying Biomedical Research Center,Lanzhou University Second Hospital,Lanzhou 730030,China;Department of Rheumatology and Immunology,Lanzhou University Second Hospital,Lanzhou 730030,China)

机构地区：[1]兰州大学第二临床医学院,兰州730030 [2]兰州大学第二医院萃英生物医学研究中心,兰州730030 [3]兰州大学第二医院风湿免疫科,兰州730030

出　　处：《中华风湿病学杂志》2024年第1期50-55,I0008,共7页Chinese Journal of Rheumatology

基　　金：国家自然科学基金(82060302,82260325);兰州大学医学创新发展计划项目(lzuyxcx-2022-168);兰州大学第二医院萃英科技创新计划项目(CY2021-MS-A04,CY2021-QN-B02)。

摘　　要：目的旨在明确百日咳毒素(PTX)能否促进实验性自身免疫性肌炎(EAM)模型间质性肺疾病(ILD)的发生并探究其潜在的致病机制。方法构建单用骨骼肌匀浆或骨骼肌匀浆联合PTX诱导的EAM模型,通过组织病理学、血清学研究明确PTX与EAM小鼠ILD表型及中性粒细胞胞外诱捕网(NETs)的关系;健康小鼠单独给予PTX腹腔注射,明确是否能在体内诱发NETs形成,PTX体外干预健康人中性粒细胞明确能否诱发NETs形成。计量资料使用Shapiro-Wilk正态性检验,采用t检验或单因素方差分析,采用图基事后检验(Tukey)进一步比较组间差异。结果与单用骨骼肌匀浆组相比,使用骨骼肌匀浆联合PTX组的EAM模型肺组织有着类似于多发性肌炎/皮肌炎(PM/DM)相关ILD的多种病理学改变,以非特异型间质性肺炎、普通型间质性肺炎为主要类型;EAM组肺间质伴有明显的NETs浸润;EAM组血清NETs标记物水平明显升高,与对照组[n=5,(87±10)ng/ml]相比,肌匀浆组[n=4,(115±27)ng/ml]和EAM组[n=7,(150±50)ng/ml]循环核酸(cfDNA)水平均升高且差异具有统计学意义(F=4.24,P=0.038);与对照组(n=4,0.13±0.02)相比,肌匀浆组(n=4,0.24±0.09),EAM组(n=6,0.33±0.11)的瓜氨酸化组蛋白H3(Cit-H3)-DNA水平升高(F=6.21,P=0.016)。PTX干预后,PTX组血清cfDNA水平[n=3,(100±40)ng/ml]高于对照组[n=3,(45±12)ng/ml,t=2.27,P=0.086];PTX在体外可诱导中性粒细胞形成NETs。结论PTX可能通过诱导NETs形成促进EAM模型ILD的发生,提示EAM小鼠可以作为靶向NETs研究ILD的模型。Objective To test the hypothesis that Pertussis toxin(PTX)can promote the occurrence of interstitial lung disease(ILD)in experimental autoimmune myositis(EAM)model and clarify the potential pathogenic mechanism.Methods EAM mice model were induced by Skeletal muscle thomogenate with or without PTX,and the relationship between ILD phenotypes and neutrophil extracellular traps(NETs)infiltration was analyzed by histopathological and serological studies in EAM with PTX group and EAM without PTX group.Healthy mice were given PTX alone intraperitoneally to clarify whether NETs formation could be induced in vivo,and neutrophils separated from healthy human blood were intervened with PTX to induce NETs formation in vitro.The data was tested for normality using Shapiro-Wilk.Statistical methods and were analyzed using t-test or ANOVA,and multiple comparisons between different groups were tested using Tukey test.Results Compared with EAM without PTX group,lung tissues in EAM with PTX group had multiple pathological changes similar to polymyositis/dermatomyositis-related ILD.Nonspecific interstitial pneumonia and usual interstitial pneumonia were the main pathological types.The pulmonary interstitial lesions were accompanied by significant infiltration of NETs;and serum NETs markers levels were obviously elevated in EAM with PTX group,compared with the control group[n=5,(87±10)ng/ml],cfDNA levels were statistically significantly elevated in both the EAM without PTX group[n=4,(115±27)ng/ml]and the EAM with PTX group[n=7,(150±50)ng/ml](F=4.24,P=0.038);Cit-H3-DNA levels were elevated in the EAM without PTX group(n=4,0.24±0.09),and in the EAM EAM with PTX group(n=6,0.33±0.11)compared with the control group(n=4,0.13±0.02)(F=6.21,P=0.016).After PTX intervention,serum cfDNA levels were higher in the PTX group[n=3,(100±40)ng/ml]than in the control group[n=3,(45±12)ng/ml,t=2.27,P=0.086];PTX also induced neutrophils to form NETs in vitro.Conclusion PTX may promote the development of ILD in EAM mice model by inducing the formati

关 键 词：神经系统自身免疫性疾病 实验性 肺疾病 间质性 百日咳毒素 胞外诱捕网 

分 类 号：R593.2[医药卫生—内科学] R563[医药卫生—临床医学]